Immunosensor for detection of inhibitory neurotransmitter gamma-aminobutyric acid using quartz crystal microbalance.
A piezoelectric immunosensor for sensing the low molecular weight neurotransmitter gamma-aminobutyric acid (GABA), one of two major inhibitory neurotransmitters in the central nervous system, is described. The sensing interface consists of a dextran layer covalently attached to a self-assembled monolayer of thiolamine compound on the surface of gold electrodes of the crystals. The dextran layer is further modified with GABA molecules to act as the biosensing layer. The affinity binding of monoclonal anti-GABA antibody on the modified piezoelectric crystals is studied in real time without any additional labels. The equilibrium association constant, K(eq) for binding between anti-GABA antibody and GABA molecules is 14.5 microg x mL (-1). The detection limit for anti-GABA is approximately 10 nM. The sensitivity of the sensor at a concentration corresponding to half-maximal response is 13.6 ng/mL x Hz. The functionalized sensor substrate is subsequently used for competitive determination of different concentrations of free GABA (range of 5 microM-50 mM) in PBS-BSA buffer. The detection limit of the immunosensor for sensing GABA with maximum sensitivity is approximately 42 microM.
Wang
Tingting
T
Harrington Department of Bioengineering, Arizona State University, Tempe, Arizona 85287, USA.
Muthuswamy
Jit
J
D000162:Acoustics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D015374:Biosensing Techniques / Q000379:methods*; D002021:Buffers; D002417:Cattle; D003911:Dextrans / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D018756:GABA Antagonists / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D006046:Gold / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D008970:Molecular Weight; D011791:Quartz; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013499:Surface Properties; D005680:gamma-Aminobutyric Acid / Q000032:analysis* / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D002021:Buffers; D003911:Dextrans; D018756:GABA Antagonists; D011791:Quartz; D012710:Serum Albumin, Bovine; D005680:gamma-Aminobutyric Acid; D006046:Gold